微小RNA-133a-5p在丙泊酚防治大鼠肝脏缺血再灌注损伤中的作用及作用机制

Role of microRNA-133a-5p and its mechanism of action in prevention and treatment of hepatic ischemia-reperfusion injury in rats with propofol

目的观察微小RNA-133a-5p(miR-133a-5p)在丙泊酚防治大鼠肝脏缺血再灌注(I/R)损伤中的作用,并探讨其可能作用机制。方法①丙泊酚对I/R大鼠肝功能及肝组织miR-133a-5p、有丝分裂原激活蛋白激酶6(mitogen-activated protein kinase 6,MAPK6)mRNA表达的影响观察:取18只大鼠分为丙泊酚组、模型组及对照组,每组6只。丙泊酚组及模型组对肝脏组织进行缺血再灌注操作,丙泊酚组在缺血再灌注操作的同时注射丙泊酚0.6 mg/(kg·min)。再灌注结束时采集各组大鼠下腔静脉血4 mL,采用全自动生化分析仪检测大鼠血清肝功能指标天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT),采血后处死各组大鼠分离肝脏组织,采用qRT-PCR法检测大鼠肝组织miR-133a-5p、MAPK6 mRNA。②miR-133a-5p抑制剂联合丙泊酚对I/R大鼠肝功能及肝组织miR-133a-5p、MAPK6mRNA表达的影响观察:另取24只大鼠分为甲、乙、丙、丁组,每组6只。四组均对肝脏组织进行缺血再灌注操作,甲组在缺血再灌注操作的同时注射丙泊酚0.6 mg/(kg·min)及miR-133a-5p抑制剂,乙组、丙组在缺血再灌注操作的同时分别注射丙泊酚0.6 mg/(kg·min)、丙泊酚+等量生理盐水,丁组不做任何处理。再灌注结束时采集各组大鼠下腔静脉血4 mL,采用全自动生化分析仪检测血清ALT、AST,采血后处死各组大鼠分离肝脏组织,采用qRT-PCR法检测四组大鼠肝组织miR-133a-5p、MAPK6mRNA。结果与对照组相比,模型组大鼠血清AST、ALT活性升高,肝脏组织miR-133a-5p相对表达量降低、MAPK6mRNA相对表达量升高(P均<0.01);与模型组相比,丙泊酚组大鼠血清AST、ALT水平降低,肝组织miR-133a-5p相对表达量升高、MAPK6mRNA相对表达量降低(P均<0.05)。与乙组相比,甲组大鼠血清ALT和AST水平高,肝组织MAPK6mRNA相对表达量高(P均<0.01);与乙和丙组相比,丁组大鼠血清ALT和AST水平升高,肝组织MAPK6mRNA相对表达量高(P均<0.01)。结论注射丙泊酚后肝脏I/R大鼠的肝损伤程度减轻,肝脏组织miR-133a-5p表达升高、MAPK6 mRNA表达降低。抑制miR-133a-5p表达可逆转丙泊酚对大鼠肝脏缺血再灌注损伤的防治作用。丙泊酚可能通过促进肝脏组织miR-133a-5p表达,抑制肝组织MAPK6 mRNA表达,减轻大鼠的肝脏I/R损伤。

Objective To observe the role of microRNA-133a-5p(miR-133a-5p)in the prevention and treatment of hepatic ischemia/reperfusion(I/R)injury in rats with propofol and to investigate its mechanism.Methods①Effects of propofol on liver function and mRNA expression of miR-133a-5p and mitogen-activated protein kinase 6(MAPK6)in I/R rats:18 rats were divided into the propofol group,model group,and control group,with 6 rats in each group.Rats in the propofol group and the model group were subjected to I/R of liver tissues,and meanwhile,rats in the propofol group were injected with 0.6 mg/(kg·min)propofol.At the end of reperfusion,4 mL of blood was collected from the inferior vena ca‐va of rats in each group,and the serum liver function indexes of aspartate aminotransferase(AST)and alanine aminotrans‐ferase(ALT)were detected using an automatic biochemical analyzer.Liver tissues were separated from rats in each group by executing the rats after the collection of blood,and miR-133a-5p,and MAPK6 mRNAs of the liver tissues of the rats were detected using qRT-PCR.②Effects of miR-133a-5p inhibitor combined with propofol on liver function and expres‐sion of miR-133a-5p and MAPK6 mRNA in liver tissues of I/R rats:another 24 rats were taken and divided into groups A,B,C,and D,with 6 rats in each group.Rats in all four groups were subjected to I/R operation on liver tissues,rats in the group A were injected with 0.6 mg/(kg·min)propofol and miR-133a-5p inhibitor at the same time during the I/R opera‐tion,and meanwhile,rats in the groups B and C were injected with 0.6 mg/(kg·min)propofol,and equal volume of nor‐mal saline,and rats in the group D did not receive any treatment.At the end of reperfusion,4 mL of blood was collected from the inferior vena cava of rats in each group,and serum ALT and AST were detected using an automatic biochemical analyzer.Liver tissues were separated from rats in each group by executing them after blood collection,and miR-133a-5p and MAPK6 mRNA were detected in liver tissues of the four groups by qRT-PCR.Results Compared with the control group,rats in the model group had increased serum AST and ALT activity,decreased relative expression of miR-133a-5p and increased relative expression of MAPK6 mRNA in the liver tissues(all P<0.01);compared with the model group,rats in the propofol group had decreased serum AST and ALT levels and increased expression of miR-133a-5p in the liver tissues,and decreased MAPK6 mRNA expression(all P<0.05).Compared with group B,rats in group A had higher se‐rum ALT and AST levels and higher relative expression of MAPK6 mRNA in the liver tissues(all P<0.01);compared with groups B and C,rats in the group D had increased serum ALT and AST levels and higher relative expression of MAPK6 mRNA in the liver tissues(all P<0.01).Conclusions The degree of liver injury is reduced in hepatic I/R rats after pro‐pofol injection,and miR-133a-5p expression increases and MAPK6 mRNA expression decreases in the liver tissues.Inhi‐bition of miR-133a-5p expression reverses the preventive effect of propofol on hepatic I/R injury in rats.Propofol may at‐tenuate hepatic I/R injury in rats by promoting miR-133a-5p expression and inhibiting MAPK6 mRNA expression in the liver tissues.